Cell-Derived Lineage Following g-Radiation-Induced Gut Injury in Mice ... âKlf4 of sham and irradiated mice day 6 after tamoxifen administration. A total of 10 ...
Stem Cell Reports, Volume 6
Supplemental Information
Krüppel-like Factor 4 Modulates Development of BMI1+ Intestinal Stem Cell-Derived Lineage Following g-Radiation-Induced Gut Injury in Mice Jes G. Kuruvilla, Chang-Kyung Kim, Amr M. Ghaleb, Agnieszka B. Bialkowska, Calvin J. Kuo, and Vincent W. Yang
Stem Cell Reports
Supplemental Information Containing Supplemental Figures, Supplemental Figure Legends, and Supplemental Experimental Procedures
Krüppel-like factor 4 modulates development of BMI1+ intestinal stem cell-derived lineage following radiation-induced gut injury in mice Jes G. Kuruvilla, Chang-Kyung Kim, Amr M. Ghaleb, Agnieszka B. Bialkowska, Calvin J. Kuo, and Vincent W. Yang
SUPPLEMENTAL FIGURE LEGENDS
Figure S1, related to Figures 1-4. Schematic representation of the lineage tracing experiments. Ctrl and Bmi1Klf4 mice were injected with tamoxifen on day 0 and exposed to 12 Gy total body irradiation or sham-irradiated on day 2. Tissues were collected on days 2 and 6 from sham-irradiated mice (for lineage tracing in homeostatic conditions) or on day 6 from irradiated mice (for lineage tracing following radiation). EdU was administered 4 hr prior to euthanasia to label actively replicating cells.
Figure S2, related to Figure 4. Additional examples of lineage tracing of eYFP+ cells after irradiation. A) and B) are two additional examples of eYFP+ lineage tracing results as described in Fig. 4.
Figure S3, related to Figure 4. Quantification of apoptosis in sham and irradiated Ctrl and Bmi1Klf4 mice. Ctrl and Bmi1Klf4 mice were injected with tamoxifen and subjected to 12 Gy total body irradiation on day 2 post-injection. Intestinal tissues were collected on day 6 post-injection (day 4 post-irradiation). A) Immunofluorescence staining of eYFP (green) and TUNEL staining as a maker for apoptosis (red) in the intestines of sham and irradiated Ctrl and Bmi1Klf4 mice on day 6 was performed. Shown is a representative figure. Panels a-d represent co-localization of TUNEL-positive cells in eYFP+ crypts. Panels e-h represent magnified views of the boxed areas in a-d. Arrowheads point to positive TUNEL staining in the respective eYFP+ crypts. B) Quantification of the number of TUNEL-positive cells in crypts containing eYFP+ cells in Ctrl and Bmi1Klf4 of sham and irradiated mice day 6 after tamoxifen administration. A total of 10 crypts containing eYFP+ cells were counted per mouse (N=3 per group). Statistical analysis was performed using a linear mixed model for repeated measurements with SAS 9.3 (SAS Institute Inc., Cary, NC). ***p
